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Reference genes for qPCR expression in black tiger shrimp, Penaeus monodon
Preety Sweta Hembrom1, Mottakunja Deepthi1, Gourav Biswas1
1Department of Genomic Science, School of Biological Sciences, Central University of Kerala, Kasaragod, Kerala, 671316, India.
Molecular Biology Reports
|March 15, 2024
Summary
This study identifies the most stable reference genes for quantitative PCR (qPCR) in Black Tiger Shrimp (P. monodon) growth research. The 18S and CLTC genes are recommended for accurate gene expression analysis in this important aquaculture species.
Area of Science:
- Aquaculture
- Molecular Biology
- Marine Biology
Background:
- Quantitative PCR (qPCR) is crucial for understanding growth and development mechanisms.
- Accurate reference gene selection is vital for reliable qPCR results.
- Limited data exists on reference gene stability in Black Tiger Shrimp (P. monodon) growth studies.
Purpose of the Study:
- To assess the expression stability of seven candidate reference genes in P. monodon.
- To identify the most reliable reference genes for qPCR analysis in P. monodon growth and development research.
- To provide validated reference genes for future molecular studies in P. monodon.
Main Methods:
- Evaluated seven reference genes (ACTB, 18S, EF-1α, AK, PK, cox1, CLTC) in adult P. monodon tissues (hepatopancreas, gills, stomach).
- Assessed gene expression stability using NormFinder, BestKeeper, and geNorm algorithms.
- Utilized RefFinder for comprehensive gene ranking and identified the most stable genes.
Main Results:
- 18S and CLTC were the most stable genes in hepatopancreas and stomach tissues.
- CLTC and AK demonstrated high statistical reliability in gill tissues.
- Validated the stability of selected genes using a growth-associated gene (insr-1).
Conclusions:
- 18S and CLTC are recommended as the most versatile reference genes for P. monodon growth-related qPCR studies.
- This research provides the first comprehensive assessment of reference genes for qPCR in P. monodon.
- The findings offer essential tools for advancing molecular research in P. monodon and related crustaceans.
